mouse tail fibroblast overexpressed Foxa3, Gata4 and Hnf1a
chip antibody
Foxa3
cell type
fibroblast undergoing iHep induction
overexpression
Foxa3, Gata4 and Hnf1a
time point
6d
strain
C57/B6 x 129sv
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Sonicate the chromatin in nuclear extracts to fragments by ultrasonication (Covaris S220). Primary antibodies were used for immunoprecipitation in sonication supernatant by incubation at 4℃ overnight. Protein G Dynabeads (Life Technologies, 10004D) were used for pulling down antibody-chromatin complex followed by washing and eluting. Foxa3:5 micrograms for ChIP, from Santa Cruz, cat id sc-5361. Gata4:5 micrograms for ChIP, from Santa Cruz, cat id sc-1237. Hnf1a:5 micrograms for ChIP, from Santa Cruz, cat id sc-6547. CTCF:5 micrograms for ChIP, from Active Motif, cat id 61312. H3K27ac:5 micrograms for ChIP, from Abcam, cat id ab4729. H3K27me3:5 micrograms for ChIP, from CST, cat id 9733. MinElute PCR purification kit (Qiagen) was used for ChIP fragments collection. NEBNext Ultra DNA Library Prep Kit was used for ChIP-seq library preparation.